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bioRxiv · 10.1101/147637

Highly parallel genome variant engineering with CRISPR/Cas9 in eukaryotic cells

Sadhu, M. J.·Bloom, J. S.·Day, L.·Siegel, J. J.·Kosuri, S.·Kruglyak, L.
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Abstract

Direct measurement of functional effects of DNA sequence variants throughout a genome is a major challenge. We developed a method that uses CRISPR/Cas9 to engineer many specific variants of interest in parallel in the budding yeast Saccharomyces cerevisiae, and to screen them for functional effects. We used the method to examine the functional consequences of premature termination codons (PTCs) at different locations within all annotated essential genes in yeast. We found that most PTCs were highly deleterious unless they occurred close to the C-terminal end and did not interrupt an annotated protein domain. Surprisingly, we discovered that some putatively essential genes are dispensable, while others have large dispensable regions. This approach can be used to profile the effects of large classes of variants in a high-throughput manner.

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Sadhu, M. J., Bloom, J. S., Day, L., Siegel, J. J., Kosuri, S., Kruglyak, L.. 2017-06-08. Highly parallel genome variant engineering with CRISPR/Cas9 in eukaryotic cells. https://doi.org/10.1101/147637

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